Research On The Mechanism Of Liushenqu Based On Intestinal Flora-immune Regulation In The Treatment Of Functional Dyspepsia

Objective:Previous study proved that Massa Medicata Fermentata(MMF)could promote gastric emptying in accumulating mice,reduced gastric pH,increased pepsin content and caused an increase in the expression of Lactobacillus and Bifidobacteria and a decrease of neutral bacteria such as Bacillus and Enterococci.The purpose of this study is to establish a FD model in rat to explore the efficacy of MMF in the treatment of FD and its intestinal flora-immune mechanism.Methods:56 male 10-day-old Sprqague-Dawley pups were randomly divided into diffirent groups:normal control group(Con,distilled water,n=8),a simple 0.1%iodoacetamide gavage-induced functional dyspepsia group(0.1%IA,n=8),0.1%iodoacetamide combined with water platform sleep deprivation-induced functional dyspepsia group(FD,n=8),Different dose of MMF-treated group(MMF 1g/kg,and MMF 3g/kg,n=8,respectively),positive drug group(Mosa.2mg/kg,n=8).Rats at 8 weeks old began oral administration for 21 days.After administration,the sugar water preference and gastric emptying rate of rats were detected;hematoxylin-eosin staining(HE)was used to detect the pathological changes of the duodenum of the rat;the Elisa kit was used to detect the duodenal Motilin(MTL)and serum levels of IFN-?,TNF-? and IL-6 in rats;16S rDNA amplicon sequencing was used to detect the effect of intestinal microflora in the colon contents of rats and the potential metabolism of the microflora.In addition,gas chromatography-mass spectrometry(GC/MS)was used to detect changes in Short Fatty Acids(SCFAs)in the colon contents of rats.Results:1.Model vs.Con.Sugar water preference rate of FD was significantly reduced,50.44%±17.19%(FD)vs.66.79%±14.26%(Con),respectively,P<0.05,while there was no significant difference between the 0.1%IA group and the Con group.Compared with the Con group,the gastric emptying rate of rats in the FD group was significantly lower(20.60%±13.26%vs 43.05%±6.72%,P<0.05);Histopathological results revealed that Duodenal villous structure in FD group was disordered,with obvious inflammatory cell infiltration in the lamina propria;Additionally,the level of duodenal MTL in Con was statistically different to FD,597.20±45.71 pg/mL vs.520.50±65.03 pg/mL,P<0.01.Meanwhile,there were obviously increase of Serum IFN-? and TNF-? and decrease of Serum IL-6 in Model compared to Con.The results of 16S rDNA amplicon sequencing showed that the microbial species richness of colon contents in FD group was significantly increased compared to Con.16S rDNA amplicon sequencing results showed that compared with Con group,FD group colon content microbial species richness was significantly increased.In addition,16S rDNA amplicon sequencing results showed that MMF caused changes in the structure and function of colon contents2.MMF-treated groups vs.FD.compared with the FD group,the sugar water consumption rate of rats in different dose groups of MMF(MMF 1g/kg and 3g/kg)and positive drug group(Mosa.)were significantly increased(P<0.05,P<0.05 or P<0.01 vs FD group);Gastric emptying rate in MMF 3g/kg group was significantly augment than FD group(P<0.05);Histopathological results revealed that Duodenal villous structure was disordered in MMF 3g/kg could significantly increase the length of duodenal villi epithelial cells in FD rats,improve duodenal villi atrophy and reduce the number of goblet cells;ELISA results show that compared with FD group,MMF lg/kg And 3g/kg administration group could significantly increase the duodenal MTL content of FD rats(P<0.01 or P<0.01 vs.FD group);compared with FD group,MMF lg/kg and 3g/kg-treated group can significantly reduce the serum levels of IFN-? and TNF-?(P<0.001 or P<0.001 vs.FD).In addition,16S rDNA amplicon sequencing results showed that MMF caused changes in the structure and function of the colonic flora of FD rats.Conclusion:MMF could promote the preference of sugar water and gastric emptying in FD rats,improve duodenal inflammation in FD rats,significantly increase the MTL and IL-6 content in the duodenum of rats,and significantly reduce IFN-? and TNF-? content in serum.In addition,MMF changed the structure of the colonic flora of FD rats and the content of short-chain fatty acids,indicating the mechanism of MMF on intestinal flora-immunomodulation treatment of functional dyspepsia.